Protein kinase C (PKC) is a group of phospholipid-dependent serine/threonine protein kinases, which togeth-er with protein kinase A (PKA) and protein kinase G (PKG) constitute a superfamily of serine/threonine protein kinase AGC. PKC includes classic PKC, novel PKC, atypical PKC and some members of kinase associated with PKC (PRK). PKC is wide-ly distributed in mammalian tissues and cells, which plays an important biological role in growth and metabolism, prolifera-tion and differentiation of cells. A series of studies have demonstrated that variations of multiple cells, occurrence and devel-opment of diseases are related to the abnormal expression of PKC. Therefore, designing and searching for efficient PKC inhib-itors have very important implications for synthesis of many kinds of effective drugs and treatment of a variety of clinical dis-eases including cancer, cardiovascular, and hypertension, et al. In recent years, the research on PKC inhibitors has become the focus of domestic and foreign research. A large number of literatures have reported many kinds of effective PKC inhibi-tors, and analyzed their function site, mechanism, clinical trial data and so on. The discovery of these PKC inhibitors has im-portant implications for structural analysis of PKC and the treatment of diseases. So in this paper, the efficient PKC inhibi-tors are summarized.

Objective To explore the change of serum amino acid serum in patients with liver cirrhosis and liver cancer, and its significance.Methods The serum amino acids of 46 patients with liver cirrhosis and 44 patients with liver cancer were examined with analyzer of Hitachi 835.Results In the case of patients with liver cirrhosis, the levels of glutamic acid, cystine, methionine and tyrosine in serum were significantly increased (P＜0.01),while, the level of phenylalanine in serum was obviously increased (P＜0.05). When it came to patients with liver cancer, the levels of serum threonine, tyrosine and phenylalanine were significantly increased (P＜0.01), the level of serum serine was obviously higher (P＜0.05). The level of BCAA/AAA (branched chain amino acids/aromatic amino acid) in the two groups was lower than that in the normal control group (P＜0.01). The levels of serum valine and leucine were lower than those in the normal control group (P＜0.05).Conclusion Analyzing the levels of serum amino acids has a certain significance for diagnosing and treating patients with liver cirrhosis and liver cancer.

Tumor multidrug resistance (MDR) is the leading cause of clinical chemotherapy failure and death.Researches show that the occurrence of MDR is related to P-glycoprotein,multidrug resistance-associated proteins,breast cancer resistance-associated proteins,lung resistance-related proteins and other factors.Now the reversal strategies of MDR include chemical drugs reversal,gene reversal,immune reversal,traditional Chinese medicine reversal and drug-loaded nano-system reversal,which make some progress and help to improve the effect of cancer chemotherapy.

OBJECTIVE To compare the BacT/Alert FN with Botai SN anaerobic blood culture bottles for detection of commonly encountered anaerobes.METHODS Using these two types of anaerobic bottles to culture 5 commonly encountered anaerobes in automatic blood culturing system and BacT/Alert system,and then to analyze the results.RESULTS There were all 32 anaerobic bottles reported positive results by BacT/Alert FN bottles,and only 8 positive bottles were reported by Botai SN bottles.CONCLUSIONS The performance of the BacT/Alert FN is much better than Botai SN anaerobic blood culturing bottle when it is used to detect commonly encountered anaerobes.

Objective To evaluate the clinical significance of the combined determination of serum procalcitonin (PCT),C reactive protein(CRP),prealbumin(PA)and white blood cell(WBC) in curative effect and prognosis of elderly patients with bacterial infection.Methods Serum PCT,CRP,PA and WBC levels and their dynamic changes of 70 patients with bacterial infection from intensive care unit(ICU)were measured.The patients with bacterial infection were classified into serious infection group and local infection group.And the comparison with virus infection group was made.Results The levels of PCT(18.00±3.20)μg/L and CRP(86.17±10.64)mg/L in serious infection group and local infection group[PCT(2.03±0.46)μg/L,CRP(50.24±7.45)mg/L] were increased.The contents of PA were reduced[(3.214±0.88)mg/L,(12.89± 1.03)mg/L].The levels of WBC were increased mildly[(8.89±1.23) × 109/L,(6.54±0.87) × 109/L].The differences were statistically significant between bacterial infection groups(serious infection group and local infection group) and virus infection group [PCT (0.21 ±0.06)μg/L,CRP (5.21 ±0.84) mg/L,PA (20.14 ±2.57)mg/L,WBC (5.30 ± 0.93) × 109/L] (P＜ 0.01).Serum PCT,CRP.PA and WBC showed significant differences between serious infection group and local infection group(P＜0.01).After antibiotic treatment,the levels of PCT (0.5 ± 0.08) μg/L,CRP (10.32 ± 1.65) mg/L and WBC (6.30± 0.91) × 109/L in bacterial infection group were reduced obviously and the concentration of PA (18.19±2.66)mg/L was elevated compared with before treatment [PCT(11.61±8.27)μg/L,CRP (71.80±20.09)mg/L,WBC(7.95± 1.59) × 109/L,PA(7.08±4.87)mg/L] (P＜0.01).Correlative analysis was performed between PCT and others indicators before and after antibiotic treatment in bacterial infection group and virus infection group.There was positive correlation between PCT and CRP,WBC before(r=0.586,0.445) and after treatment (r=0.688,0.463) in bacterial infection group.PCT and WBC(r=0.432) were positively correlated after treatment,while no correlation existed between PCT and others indexes in virus infection group(r＜0.306).When comparing sensitivity and specificity of PCT and CRP for diagnosis of bacterial infection disease,the sensitivity (95.7％) of PCT was similar to that of CRP.The specificity of PCT(86.7％) was remarkably higher than that of CRP(73.3％).Conclusions The combination of serum PCT,CRP.PA and WBC is valuable for early diagnosis of bacterial infection disease.Monitoring PCT level dynamically would assist an evaluation of curative effect and disease outcome of elderly patients.

Objectives This research explored the characteristics of changes in the serum metabolic profile of preeclampsia pregnancy(PE) to establish the disease distinguish model and screen characteristic metabolic markers with potential diagnostic value for preeclampsia.Methods From August 2014 to January 2016,samples in three groups were collected at Tianjin Third Central Hospital.Thirty-one clinically diagnosis patients with preeclampsia,25 normal pregnancy women and 29 healthy volunteers of childbearing age were enrolled.Ultraperformance liquid chromatography-mass spectrometry (UPLC-MS) was used to analyze serum metabolites of PE group (31 patients with preeclampsia),P group (25 normal pregnancy women) and Normal group (29 healthy volunteers of childbearing age).Nonparametric test analyzes were used to analyze the data and find the specific metabolites.Results This research established the principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) disease distinguish model for PE group,P group and Normal group.To distinguish PE group,P group and Normal group,15 characteristic metabolites were identified.Eight kinds of glycerol phospholipid (including 7 kinds of hemolysis phosphatidyl choline and 1 kind of lysophospholipids acid) and 1 kind of sphingomyelin in PE group were higher than that of normal pregnancy group.The difference had statistically significant(Z of the metabolites were 2.32,3.34,3.21,2.60,2.22,3.40,3.58,5.84,2.70 respectively,all P<0.05).1,25-Dihydroxyvitamin D3-26,23-lactone and 24-Oxo-1alpha,23,25-trihydroxyvitamin D3 in PE group were higher than that of P group and Normal group,which had a statistics difference (Z of the metabolites were 2.01,3.89,3.26,2.34 respectively,all P<0.05).Conclusions Metabolomics distinguish model has a good ability to distinguish PE group,P group and Normal group.Serum characteristic metabolites can successfully reflect the status of fat,calcium and phosphorus metabolism of preeclampsia patients and provide high value for prediction,diagnosis and treatment.

Objective To calculate the biological reference of plasma amino acid on L-8900 amino acid analyzer and to provide reference for clinical diagnosis with domestic reagents replacing original reagent .Methods By testing the original standards and the same batch of plasma (50 cases) ,we compared original reagents with domestic reagents for their performance (including resolution , repeatability ,accuracy ) .We tested the plasma free amino acids of 400 cases of healthy people using the domestic reagents to estab-lish biological reference interval of plasma amino acid ,and do correlation analysis between the amino acids level and liver function . Results (1)Domestic reagents showed high accuracy in the results of 5 consecutive detection of amino acids were high peak separa-tion and high peak retention time and high peak area .(2)Statistics derived biological reference interval of 19 amino acids and 10 kinds of amino acids had significant differences .(3)Correlation analysis showed that ALT and liver function were negatively corre-lated with threonine(P<0 .05) .GLU and valine ,isoleucine ,leucine were positively correlated(P<0 .05) .CHO and negatively cor-related with isoleucine(P<0 .05) .Conclusion Domestic agents can replace the original reagents ,on the basis ,the biological refer-ence intervals of plasma amino acids have great importance to clinical diagnosis and prognosis .

Objective To explore the level of free fatty acid (FFA)in ischemic stroke (IS) patients and its correlation with insulin resistance (IR).Methods This study was case-control study.Patients who were diagnosed with IS were followed up from June,2011 to September,2012 in Tianjin Third Central Hospital.The 180 IS patients were divided into large infarct(57),middle infarct(63)and small infarct (60) according to the area of infarct.At the same time,60 healthy persons were selected as control group.The plasma FFA,fasting insulin (FINS),fasting blood glucose (FBG),homeostasis model assessment (HOMA)-IR,triacylglycerol (TG),total cholesterol (TC),high density lipoprotein cholesterol-C (HDL),low density lipoprotein cholesterol-C(LDL),lipoprotein A1 (APOA1) and lipoprotein B (APOB) in 180 IS patients were measured and compared with that of control group.The correlation between FFA and IR was analyzed by using pearson linear correlation.The relationship between FFA and HOMA-IR was analyzed by using multiple stepwise multivariate analysis.The t test and analysis of variance were used for statistical analysis.Results Compared to normal control,the level of FFA,FINS,FBG,FIB,TG,TC,LDL,APOB and HOMA-IR in IS patients were higher,the difference was statistically.The level of HDL,APOA1 in IS patients were lower than that of normal control.The level of FFA,FINS,HOMA-IR,FBG,TG,TC,HDL,LDL,APOA1 and APOB in IS patients related to the area of infarct.The level of FFA,FINS,HOMA-IR,FBG,TG,TC,LDL,APOB in large infarct were higher than those in middle infarct and small infarct.The level of HDL,APOA1 in large infarct were lower than those in middle infarct and small infarct,the difference were statistically.In IS patients,FFA had positive correlation to FINS,FBG and HOMA-IR (r =0.870,0.497,0.792,P ＜ 0.001),but had no correlation to age (r =0.008,P ＞ 0.05).In IS patients,HOMA-IR was dependent variable,but FFA,FINS,FBG,TG,TC,HDL,APOA1 and APOB were independent variable.FFA was independent risk factor to HOMA-IR,besides FINS,FBG,TC,LDL and APOB.Conclusion FFA elevates in IS patients,FFA has correlation to IR and it is a dependent risk factor to IS patients.

Abstract: Cell death is a fundamental biological phenomenon that is essential for the survival and development of organisms. Cell death can be either a spontaneous programmed process by the host or an accidentally triggered process. According to the different signaling pathway activated by various stimulates, programmed cell death exhibits the lytic or non-lytic morphology. For example, apoptosis, a typical non-lytic form of cell death, exhibits cell shrinkage and induces the formation of apoptotic bodies. Pyroptosis mediated by cysteine-containing aspartate-specific protease-1/11 (caspase-1/11) and necroptosis can induce inflammatory reactions and promote cell lysis to release inflammatory cytokines via triggering the pore-forming mechanism of the cell membrane, representing a typical modes of lytic cell death. In addition, the release of reactive oxygen species caused by the damaged mitochondria may further trigger ferroptosis during the pathogen infection. Programmed cell death can play an immune defensive role by eliminating infected cells and intracellular pathogens and stimulating the innate immune response through the resulting cell corpses. Here, we discuss the molecular mechanisms of five programmed cell death pathways: apoptosis, pyroptosis, ferroptosis, necroptosis and PANoptosis. We describe their roles in the innate immune defense against bacterial infections and give a brief statement of the interactions between the different programmed cell death, hoping to provide new insights for in-depth study of the pathogenic mechanisms of infectious diseases.

BACKGROUND: The skill to culture osteoblasts primarily has been well developed. However, trypsinase can affect membrane protein of osteoblasts if the time of digestion is long. Therefore, it is of great significance to select an ideal method to avoid the damage from trypsinase to cells as possible when culturing osteoblasts.OBJECTIVE: To explore a novel method to isolate and culture SD rat osteoblasts in vitro, and identify the functions of the cells.DESIGN: Observational study.SETTING: Department of Orthopaedics, First Affiliated Hospital of Jinan University.MATERIALS: This experiment was carried out in the Department of Orthopaedics, First Affiliated Hospital of Jinan University from March to May in 2007. Eight SPF 24-hour old SD rats were used in the experiment. The rats, irrespective of gender, were provided by the Experimental Animal Center of Nanfang Medical University. The experimental animals were disposed according to ethical criteria. The main reagents were detailed as follows: collagenase Ⅱ (Sigma Company);trypsin (Sigma Company); alkaline phosphatase (ALP) kit (Nanjing Jiancheng Biological Products Company); SABC-1021(Wuhan Boster Biotechnology Company).METHODS: 24-hour old SD rats were chosen for experiment. The newly born SD rats were sacrificed by anesthesia and the cranial bones of the rats were obtained cleanly, erased completely of the periosteum and cut to blocks of I mm3. The cranial bones were digested by 0.25 % trypsinase for 20 minutes, then by 0. 1% type Ⅱ collagenase for 60 minutes. The digestive time of trypsinase was controlled in the process of digestion to avoid to harm the cells. The liquid was gathered and centrifuged. The cells were cultured in culture flask and were purified by many times adhered.MAIN OUTCOME MEASURES: Morphology observations under the inverted phase contrast microscope, transmission electron microscope, and scanning electron microscope were performed. The phenotype, calcium tuberculation and the expression of alkaline phosphatase were studied with alizarin red staining and modified Gomori Ca-Co assays respectively.The cells were also evaluated with collage Ⅰ immunohistochemical staining.RESULTS: The cultured cells had active proliferation ability. Cells showed multi-angle or fusiform shape. Nucleus was immature and organell was plentiful. Therefore, they had typical morphological characters of osteoblasts. Moreover, they showed the osteoblastic phenotypes such as their synthesis of alkaline phosphatase, collage Ⅰ and formation of calcium tuberculations.CONCLUSION: The cells cultured by our modified enzymatic digestion method had typical morphological and biological characteristics of osteoblasts.